在p13Suc1上对有丝分裂后期促进复合物/细胞周期体进行亲和纯化。
Affinity purification of mitotic anaphase-promoting complex/cyclosome on p13Suc1.
作者信息
Hershko Avram
机构信息
Unit of Biochemistry, Technion-Israel Institute of Technology, Haifa 31096, Israel.
出版信息
Methods Enzymol. 2005;398:170-5. doi: 10.1016/S0076-6879(05)98015-4.
A procedure is described for the affinity purification of the mitotic form of anaphase-promoting complex/cyclosome (APC/C) from HeLa cells. It is based on the binding of mitotically phosphorylated APC/C to the phosphate-binding site of p13(suc1), followed by specific elution with a phosphate-containing compound. The procedure is rapid, simple, and yields 50- to 70-fold purification of soluble APC/C, with a approximately 30% recovery of activity.
本文描述了一种从HeLa细胞中亲和纯化后期促进复合物/细胞周期体（APC/C）有丝分裂形式的方法。该方法基于有丝分裂磷酸化的APC/C与p13（suc1）的磷酸结合位点的结合，随后用含磷酸盐的化合物进行特异性洗脱。该方法快速、简单，可使可溶性APC/C得到50至70倍的纯化，活性回收率约为30%。
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